Herpes simplex viruses (HSV) are common human infections which cause a wide spectrum of diseases ranging from benign fever blisters to life- threatening HSV encephalitis (HSE) and neonatal HSV infections. Significant therapeutic advances have been achieved with drugs such as acyclovir, particularly for mucocutaneous infections. Neonatal HSV infections and HSE are still associated with significant morbidity and mortality. If treatment is to be advanced for life-threatening diseases, the development of rapid diagnostic technology is essential for early diagnosis and to identify individuals at high risk for severe disease. The proposed studies represent a collaborative effort between two highly interactive groups at the University of Alabama at Birmingham and The University of Chicago interested in applying the fruits of research on the molecular biology of HSV to clinical research. We have established three objectives. The aims of Objective 1 are to develop reagents for determining past exposure of individuals to HSV-1 and HSV-2, whether by natural infection or vaccination. The emphasis of the experimental design is on type-specific peptides as reagents for serologic evaluations. The reliability of these assays will be compared with those on glycoproteins (g) G1 and G2. Objective 2 will establish methods for very rapid detection of viral nucleic acids in biological specimens such as CSF, vaginal washes, and tissue specimens. We will compare results of our established immunoblot ELISA assay and a unique combined PCR-liquid phase DNA hybridization assay with routine virus isolation. Objective 3 applies the methods of Objectives 1 and 2 to prospective studies of excretion and transmission of HSV from pregnant women to the newborn at delivery and CSF specimens obtained from patients with biopsy-proven HSE, disease of suspected HSV etiology, and CNS disease of unknown etiology. Consistent with the structure of this collaborative effort, the success of a research proposal such as this one is dependent upon (i) the availability of clinical specimens that have been precisely classified according to clinical history, viral isolation and typing, and type-specific serologic statu by immunoblot and gG1 and gG2 and (II) the ability to generate and apply suitable molecular probes. We have in our repositories catalogued biologic specimens for these studies including serially collected serum and genital swabs from pregnant women and students with primary genital herpes, and CSF, serum, and tissue from patients with neonatal HSV and HSE.